Anti-inflammatory method

ABSTRACT

The invention relates to the use of carbocyclically and heterocyclically fused dihydropyridines as cerebroprotective agents, as agents for treating chronic inflammatory processes and as agents for inhibiting blood clotting, and also relates to new compounds of formula Ie ##STR1##

This is a division of application Ser. No. 08/238,298, filed May 5, 1994now abandoned, which is a continuation of application Ser. No.08/036,299, filed Mar. 24, 1993 (abandoned), which is a continuation ofapplication Ser. No. 07/812,321, filed Dec. 23, 1991 (abandoned), whichis a continuation of application Ser. No. 07/809,610, filed Dec. 17,1991 (abandoned).

Dihydroisoquinolines of general formula Ia are known from EP-A 37 934.The compounds mentioned therein have a cardiotonic effect and have theeffect of increasing contractility and influencing blood pressure. Theyhave been proposed for improving the blood flow through the tissues andimproving the oxygen supply to the tissues. These possible applicationsare based on the vascular activity of the compounds. EP-A 251 194describes how carbocyclically and heterocyclically fuseddihydropyridines have a cardioprotective effect and constitute a totallynew type of Ca-antagonistic compounds.

The present invention relates to the use of the compounds known from theabove-mentioned EP-A 251 194 as cerebroprotective agents, particularlyfor treating patients who have suffered a stroke or are at risk ofsuffering a stroke. The invention further relates to the use of thesecompounds for treating chronic inflammatory processes (e.g. bronchialasthma, arthritis) and for inhibiting blood coagulation or bloodplatelet aggregation. The invention further relates to a group of newfused dihydropyridines, namely new 3,4-dihydroisoquinoline derivativesof general formula Ie as defined hereinafter.

The invention thus relates to the use of carbocyclically andheterocyclically fused dihydropyridines of the formula ##STR2##including the tautomeric forms thereof of formula II, this termrepresenting the cis- and trans-forms II' and II" together: ##STR3##wherein X represents OR₁, NHR₂ ; NR₃ R₄ and

R₁ represents hydrogen, C₁₋₄ -alkyl, C₁₋₄ -alkoxyalkyl

R₂ represents hydrogen; C₃₋₆ -alkenyl, C₃₋₆ -alkynyl; C₃₋₆ -cycloalkyl;C₃₋₆ -cycloalkenyl; straight-chained or branched C₁₋₆ -alkyl which mayif desired be mono- or polysubstituted with the substituents of groupsa) to c) listed below, which may be identical or different:

a) halogen; cyano; hydroxy; mercapto; C₁₋₄ -alkoxy; C₁₋₄ -alkylthio;amino; mono-C₁₋₄ -alkylamino; di-C₁₋₄ -alkylamino (wherein the alkylradical may be identical or different), phenoxy (wherein the phenylgroup may be substituted as in b) below)

b) phenyl; optionally mono- or polysubstituted, by identical ordifferent substituents, by the groups halogen, trifluoromethyl C₁₋₄-alkoxy, hydroxy, mercapto, C₁₋₄ -alkylthio, C₁₋₄ -alkyl, amino,mono-C₁₋₄ -alkylamino, di-C₁₋₄ -alkylamino (wherein the alkyl groups maybe identical or different), C₂₋₃ -acylamino, C₂₋₃ -acyloxy and the group--O--(CH₂)_(n) --O vicinally by bound to the phenyl system (wherein n=1or 2)

c) a 5- or 6-membered saturated or wholly or partially unsaturatedmonocyclic heterocyclic group having up to 3 heteroatoms selected fromthe group N, O, S; and as a bicyclic heterocycle indole (whilst theabove-mentioned heterocycles may be mono- or polysubstituted by C₁₋₄-alkyl), C₃₋₆ -cycloalkyl; C₅ - or C₆ -cycloalkenyl; C₂₋₃ -acyl; C₁₋₄-alkylsulphonyl; or phenyl (which may in turn be substituted up to threetimes as described in b));

or R₂ represents phenyl, which may be substituted as described in b)above;

R₃ and R₄ independently of each other represent C₁₋₄ -alkyl, which mayif desired be phenyl-substituted, whilst the phenyl substituent may inturn be substituted as under b) hereinbefore;

or

R₃ and R₄ together with the nitrogen atom to which they are boundrepresent a wholly or partially saturated heterocyclic 5- or 6-memberedring (which may also contain up to 2 further heteroatoms from the groupN, O, S), whilst the heterocyclic group thus obtained may be substitutedby C₁₋₄ -alkyl, hydroxy or (CH₂)_(p) --R₅ (where p=0 or 1)

and

R₅ represents a phenyl radical which is optionally substituted as underb) hereinbefore;

A represents the fused ring systems ##STR4##

wherein

R₈ represents hydrogen; C₁₋₄ -alkyl or C₁₋₄ -alkoxy

R₆ and R₇ which may be identical or different represent hydrogen;hydroxy; C₁₋₄ -alkyl; C₁₋₄ -alkoxy; amino; methanesulphonylamino

or

R₆ and R₇ together represent --O--(CH₂)_(n) --O-- (where n=1 or 2)

R₉ represents hydrogen; C₁₋₄ -alkyl

R₁₀ represents hydrogen or 2-phenyl-2-ethoxycarbonylacetyl

and the salts thereof with physiologically acceptable acids, bases orcomplex-forming agents for cerebral protection, for treating chronicinflammatory processes and for inhibiting blood coagulation or bloodplatelet aggregation.

Within the general formula I are included the new,3,4-dihydroisoquinoline derivatives of general formula Ie ##STR5##wherein X represents a group ##STR6## in which X₁ represents a phenylgroup, mono- or di-substituted by a trifluoromethyl or ethoxy group, aphenyl group or a 2-methoxyphenyl group substituted by a fluorine atomand a methoxy group;

X₂ represents a group --CH₂ --CH₂ -- or CH₂ --CH(CH₃); and

X₃ represents a 2,3,4-trimethoxyphenyl, 2,3-dimethoxyphenyl,2,6-dimethoxyphenyl or 3,6-dimethoxyphenyl group, mono- ordi-substituted by a trifluoromethyl or ethoxy group or a phenyl groupsubstituted by a methoxy group and a fluorine atom;

and their pharmaceutically acceptable salts.

These new compounds are also suitable for the uses described above.

Preferred are compounds of general formula Ie wherein X represents agroup ##STR7## in which X₁ represents a 2-methoxyphenyl group which mayoptionally be additionally substituted by fluorine atoms;

X₂ represents a --CH₂ CH₂ -- group; and

X₃ represents a 2,3,4-trimethoxyphenyl, 2,3-dimethoxyphenyl,2,6-dimethoxyphenyl, 3,6-dimethoxyphenyl, a 2- or 3-thienyl group, aphenyl group substituted by a trifluoromethyl or ethoxy group or aphenyl group substituted by a methoxy group and a fluorine atom.

It is preferred to use carbocyclically and heterocyclically fuseddihydropyridines of formula ##STR8## and the tautomeric forms thereof offormula ##STR9## wherein X represents OR₁ NHR₂ ; NR₃ R₄ R₁ representsmethyl or ethyl

R₂ represents hydrogen; straight-chained or branched unsubstituted C₁₋₅-alkyl; allyl; propargyl; C₃₋₆ -cycloalkyl; 3-chlorophenyl;2-methyl-3-chlorophenyl; or C₁₋₃ -alkyl, which is mono-substituted withone of the substituents of groups d) to f) listed hereinafter;

d) cyano, hydroxy, methoxy, dimethylamino

e) phenyl, 3,4-methylenedioxyphenyl, mono-, di- or trimethoxysubstituted phenyl, 3-methoxy-4-hydroxyphenyl,3-hydroxy-4-methoxyphenyl,

f) morpholino, pyridin-2-yl, indol-3-yl, furan-2-yl, thiphen-2-yl,pyridin-3-yl, pyridin-4-yl

R₃ and R₄ independently of each other represent methyl; ethyl;3-cyanopropyl; benzyl; or 3,4,5-trimethoxyphenethyl or

R₃ and R₄ together with the nitrogen atom to which they are boundrepresent morpholine; thiomorpholine; pyrrolidine; piperazine;4-methylpiperazine; 4-benzylpiperazine; or4-(2-methoxyphenyl)piperazine;

and

A represents the fused ring systems ##STR10##

wherein

R₈ represents hydrogen or methoxy

R₆ represents methoxy; hydroxy; hydrogen; amino; ormethanesulphonylamino

R₇ represents hydrogen; methoxy; or hydroxy

R₉ represents methyl

R₁₀ represents 2-phenyl-2-ethoxycarbonylacetyl; or hydrogen

and the salts thereof with physiologically acceptable acids.

The invention thus relates to dihydroisoquinolines of the formula##STR11## dihydro-thieno 3,2-c!pyridines of the formula ##STR12##dihydro-pyrrolo 3,2-c!pyridines of formula ##STR13## and dihydro-pyrido3,4-b!indoles of the formula ##STR14## and the tautomeric forms thereofIIa', IIb', IIc' and IId' and the forms IIa", IIb", IIc" and IId" whichare E/Z-isomeric thereto.

The compounds of formulae Ia to Id are chiral. The invention alsoincludes the two R- and S-enantiomeric forms of the compounds of formulaI wherein the meanings of the groups X, R₆, R₇, R₈, R₉ and R₁₀ aredefined as hereinbefore.

The substances listed in Tables 1 to 12 hereinafter preferably occur inthe tautomeric form I or II. In Table 12 the new compounds of generalformula Ie are listed.

The preferred tautomeric form is designated I or II in the "structure"column.

The following compounds of formula I in tautomeric form I or II arelisted by name.

                  TABLE 1                                                         ______________________________________                                        Structural type:                                                               ##STR15##                                                                    No.     X                      Structure                                      ______________________________________                                         1.     OCH.sub.3              II                                              2.     OC.sub.2 H.sub.5       II                                              3.     NHCH.sub.3             I                                               4.     NHC.sub.2 H.sub.5      I                                               5.     NH(CH.sub.2).sub.2CH.sub.3                                                                           I                                               6.     NH(CH.sub.2).sub.3CH.sub.3                                                                           I                                               7.     NH(CH.sub.2).sub.4CH.sub.3                                                                           I                                               8.     NHCH(CH.sub.3).sub.2   I                                               9.     NHCH.sub.2CH(CH.sub.3).sub.2                                                                         I                                              10.     NH(CH.sub.2).sub.2CH(CH.sub.3).sub.2                                                                 I                                              11.     NHC(CH.sub.3).sub.3    I                                              12.     NHCH(CH.sub.3)C.sub.2 H.sub.5                                                                        I                                              13.     NHCH.sub.2CHCH.sub.2   I                                              14.     NHCH.sub.2CCH          I                                              15.     NH(CH.sub.2).sub.2OH   II                                             16.     NHCH.sub.2CH(OH)CH.sub.3                                                                             I                                              17.     NH(CH.sub.2).sub.2OCH.sub.3                                                                          II                                             18.     NH(CH.sub.2).sub.3OCH.sub.3                                                                          II                                             19.     NH(CH.sub.2).sub.2N(CH.sub.3).sub.2                                                                  I                                              20.     NH(CH.sub.2).sub.3N(CH.sub.3).sub.2                                                                  II                                                      ##STR16##             II                                                      ##STR17##             I                                                       ##STR18##             I                                                       ##STR19##             I                                                       ##STR20##             I                                                       ##STR21##             II                                                      ##STR22##             II                                                      ##STR23##             I                                                       ##STR24##             I                                              30.                                                                                    ##STR25##             II                                                      ##STR26##             I                                                       ##STR27##             II                                                      ##STR28##             I                                                       ##STR29##             I                                                       ##STR30##             II                                             36.     N(CH.sub.3).sub.2      I                                              37.     N(C.sub.2 H.sub.5).sub.2                                                                             I                                                       ##STR31##             I                                              38a.                                                                                   ##STR32##             I                                                       ##STR33##             I                                              40.                                                                                    ##STR34##             I                                                       ##STR35##             I II                                                    ##STR36##             II                                                      ##STR37##             I                                                       ##STR38##             I                                              ______________________________________                                    

                  TABLE 2                                                         ______________________________________                                        Structural type:                                                               ##STR39##                                                                    No.     X                     Structure                                       ______________________________________                                        45.     OC.sub.2 H.sub.5      II                                                       ##STR40##            II                                                       ##STR41##            II                                                       ##STR42##            II                                                       ##STR43##            II                                              50.                                                                                    ##STR44##            I                                               ______________________________________                                    

                  TABLE 3                                                         ______________________________________                                        Structural type:                                                               ##STR45##                                                                    No.     X                    Structure                                        ______________________________________                                        51.     OC.sub.2 H.sub.5     I                                                                             II                                               52.     NH(CH.sub.2).sub.3CH.sub.3                                                                         I                                                53.     NH(CH.sub.2).sub.4CH.sub.3                                                                         I                                                54.     NHCH(CH.sub.3).sub.2 I                                                55.     NHCH.sub.2CH(CH.sub.3).sub.2                                                                       I                                                56.     NHCH.sub.2 CHCH.sub.2                                                                              I                                                         ##STR46##           I                                                         ##STR47##           I                                                         ##STR48##           I                                                60.                                                                                    ##STR49##           I                                                         ##STR50##           I                                                         ##STR51##           I                                                         ##STR52##           I                                                         ##STR53##           I                                                         ##STR54##           I                                                         ##STR55##           I                                                         ##STR56##           I                                                         ##STR57##           I                                                ______________________________________                                    

                  TABLE 4                                                         ______________________________________                                        Structural type:                                                               ##STR58##                                                                    No.           X        Structure                                              ______________________________________                                        69.           N(C.sub.2 H.sub.5).sub.2                                                               I                                                      ______________________________________                                    

                  TABLE 5                                                         ______________________________________                                        Structural type:                                                               ##STR59##                                                                    No.       X                Structure                                          ______________________________________                                        70.       NHCH.sub.3       I                                                  71.       NHC.sub.2 H.sub.5                                                                              I                                                             ##STR60##       I                                                             ##STR61##       I                                                  74.       N(CH.sub.3)C.sub.2 H.sub.5                                                                     I                                                  ______________________________________                                    

                  TABLE 6                                                         ______________________________________                                        Structural type:                                                               ##STR62##                                                                    No.     X                  Structure                                          ______________________________________                                                 ##STR63##         I                                                           ##STR64##         I                                                  77.     N(CH.sub.3)C.sub.2 H.sub.5                                                                       I                                                           ##STR65##         II                                                          ##STR66##         I                                                  ______________________________________                                    

                  TABLE 7                                                         ______________________________________                                        Structural type:                                                               ##STR67##                                                                    No.           X        Structure                                              ______________________________________                                        80.           OC.sub.2 H.sub.5                                                                       II                                                     ______________________________________                                    

                  TABLE 8                                                         ______________________________________                                        Structural type:                                                               ##STR68##                                                                    No.           X        Structure                                              ______________________________________                                        81.           OC.sub.2 H.sub.5                                                                       II                                                     ______________________________________                                    

                  TABLE 9                                                         ______________________________________                                        Structural type:                                                               ##STR69##                                                                    No.   X                       Structure                                       ______________________________________                                        82.   OC.sub.2 H.sub.5        I                                                                             II                                                     ##STR70##              II                                                     ##STR71##              II                                                     ##STR72##              I II                                                   ##STR73##              I                                               87.   NHCH.sub.2CH(CH.sub.3).sub.2                                                                          II                                              88.   NH(CH.sub.2).sub.3N(CH.sub.3).sub.2                                                                   II                                                     ##STR74##              II                                              ______________________________________                                    

                  TABLE 10                                                        ______________________________________                                        Structural type:                                                               ##STR75##                                                                    No.       X                Structure                                          ______________________________________                                        90.       OC.sub.2 H.sub.5                                                               ##STR76##       II                                                 ______________________________________                                    

                  TABLE 11                                                        ______________________________________                                        Structural type:                                                               ##STR77##                                                                    No.       X                Structure                                          ______________________________________                                        92.       OC.sub.2 H.sub.5 II                                                 ______________________________________                                    

                                      TABLE 12                                    __________________________________________________________________________    Structural type:                                                               ##STR78##                                                                    II                                                                            Compound                                                                             X                          Structure                                                                            M.pt(°C.)                     __________________________________________________________________________            ##STR79##                 I      56-64                                B                                                                                     ##STR80##                 I      176-184                              C                                                                                     ##STR81##                 I      166-168                              D                                                                                     ##STR82##                 II     102-104                              E                                                                                     ##STR83##                 I      187                                  F                                                                                     ##STR84##                 I      94-96                                G                                                                                     ##STR85##                 II     139-142                              H                                                                                     ##STR86##                 II     133-135                              J                                                                                     ##STR87##                 II     143-145                              K                                                                                     ##STR88##                 II     96-99                                L                                                                                     ##STR89##                 II     118-120                              M                                                                                     ##STR90##                 II     122-114                              N                                                                                     ##STR91##                 I      95-99                                O                                                                                     ##STR92##                 I      114-116                              P                                                                                     ##STR93##                 I      66-73                                Q                                                                                     ##STR94##                 II     205-209                              __________________________________________________________________________

In the definitions used in the text the radicals and groups may beidentical or different, i.e. if one of the above-mentioned substituentsoccurs several times in a particular molecule, the meaning can beselected freely within the scope of the definitions provided.

The term alkyl means C₁₋₆ -alkyl and C₁₋₄ -alkyl radicals which may besubstituted or, as alkyl radicals, are part of a functional group suchas alkoxy or alkylthio. The alkyl radicals include methyl, ethyl,n-propyl, isopropyl, n-butyl, sec.-butyl, isobutyl and tert.-butylradicals as well as the various isomeric pentyl and hexyl radicals, suchas e.g. isopentyl, neopentyl, n-pentyl and n-hexyl radicals.

The above definition thus also applies even when the alkyl radicalitself is substituted and/or is itself part of an alkoxyalkyl,alkoxycarbonyl, alkoxy, alkylthio, alkylsulphonyl, monoalkylamino,alkylmethyl, alkylthiomethyl or dialkylamino group or the alkyl radical,as a substituent, is bound to an aromatic heterocyclic or carbocyclicsystem.

The halogens are fluorine, chlorine, bromine and iodine, preferablyfluorine, chlorine and bromine and, to a lesser extent, iodine.

C₃₋₆ -cycloalkyl indicates cyclopropane, cyclobutane, cyclopentane andcyclohexane.

C₅₋₆ -cycloalkenes denote e.g. cyclopentene, cyclohexene andcyclohexadiene.

The C₂ - and C₃ -acyl radicals denote acetyl and propionyl radicals.

C₃₋₆ -alkynes are the isomeric hexynes, pentynes, butynes and propynes,preferably propargyl.

The C₃₋₆ -alkenes are the isomeric hexenes, pentenes, butenes andpropenes, preferably allyl.

Examples of unsaturated heterocyclic groups include, inter alia:

furan, pyran, pyrrole, pyrazole, imidazole, pyridine, pyrazine,pyrimidine, pyridazine, thiophene, thiazole, oxazole, 1,2,4-triazole,1,2,3-triazole, 1,2,4-triazine, 1,3,5-triazine, indole.

Examples of 5- or 6-membered, wholly or partially saturated monocyclicheterocycles include, inter alia:

imidazolidine, pyrazolidine, pyrrolidine, piperidine, piperazine,morpholine, thiomorpholine, tetrahydrofuran, tetrahydrothiophene,1,4-dioxine, imidazoline, pyrazoline, pyrroline, etc.

The compounds of formula I or II are bases and can be converted in theusual way with inorganic or organic acids and salts and complex-formingagents into any desired physiologically acceptable adducts (salts).

Acids suitable for salt formation include for example hydrochloric,hydrobromic, hydriodic, hydrofluoric, sulphuric, phosphoric, nitric,acetic, propionic, butyric, caproic, valeric, oxalic, malonic, succinic,maleic, fumaric, lactic, tartaric, citric, malic, benzoic,p-hydroxybenzoic, phthalic, cinnamic, salicylic, ascorbic,methanesulphonic acid and the like.

The majority of the compounds of general formula I and the methods ofpreparation thereof are known from EP-A 37 934 and EP-A 251 194 referredto above. These publications do not mention the compounds of formula Ie.

As already mentioned hereinbefore, the subject of the present inventionis the use of the compounds known from the above-mentioned EP-A 251 194and particularly the new compound of Example 38A) as a cerebroprotectiveagent. The compounds are valuable in the treatment of degenerative andnecrotic diseases of the brain. It is also possible to providepreventative treatment for patients who are at risk from such diseases.As is shown by the experiments described hereinafter, the effect of thecompounds is not based on an improvement in the blood flow through thetissues. The compounds are therefore suitable for a new kind oftreatment of epilepsy and Alzheimer's disease and particularly fortreating patients who have suffered a stroke or are at risk of sufferinga stroke. Moreover, as mentioned earlier, the compounds are suitable fortreating chronic inflammatory processes and for inhibiting bloodclotting.

The following test results show the surprising efficacy of thecompounds. The tests were carried out particularly on Compound A(Example 38A) ##STR95## as a representative example of the compounds ofgeneral formula I. Ischaemia tolerance in the gerbil: (e.g. Suzuki, R.et al., Acta Neuropath (Berl.) 1983, 60:207-216, 217-222; Yoshidomi, M.et al., J. Neurochem. 1989, 53:1589-1594) Ischaemia caused by occlusionof the carotid artery for 10 minutes under ether anaesthesia. Compound Awas administered in doses of 1 and 10 mg/kg s.c. 4×within 24 hours,beginning 2 hours after the reopening of the arteries.

72 hours after the occlusion of the arteries the animals were killed andthe brains were dissected for histological examination. As evidence ofischaemic damage or a reduction thereof, the damage to the cells in thevicinity of the CA₁ -region of the hippocampus in a defined section ofthe histological preparation was evaluated. The test was carried out ongroups of 5 animals.

In the group in which the animals were not given a test compound, allthe animals showed significant damage to the CA₁ -region examined. Bycontrast, when Compound A was administered, there was a cleardosage-dependent protection against the damage of hypoxia. When 1 mg/kgof Compound A was administered, only 2 out of 5 animals had any damageto the CA₁ -region. When 10 mg/kg of Compound A were administered, noneof the 5 animals showed any damage.

These results indicate that the compounds of general formula I can beused for the curative treatment of neurological damage, e.g. as causedby strokes.

The results of this trial also show that the compounds break through theblood-brain barrier, which is a major feature of the invention.

The following tests on isolated cell cultures show the mechanism ofactivity of the test compounds. From these test results (as from theresults of tests on gerbils) it can be concluded that the compounds ofgeneral formula I can be used for the treatments mentioned in thispatent application.

In isolated cell cultures (e.g. neutrophilic granulocytes, HL 60-cells,thrombocytes, etc.) Compound A was able to inhibit the cell death and"calcium overload" provoked by different agonists (e.g. PAF,leucotrienes, endothelin, FMLP or isoprenalin) as a function of thedosage (IC₅₀ values in the region of 1 μM).

The half-maximum inhibitory concentrations of test substances whichinhibit the FMLP (1 nM)-induced calcium transients on FLUO₃ -chargedHL60 cells were measured.

    ______________________________________                                        Compound                                                                      No.          Half-maximum inhibitory concentration                            ______________________________________                                        43             5.10.sup.-5 M                                                  47           4,8.10.sup.-5 M                                                  38a (Compound A)                                                                           5,4.10.sup.-5 M                                                  ______________________________________                                    

The test methods mentioned above are described by W. K. Pollock. T. J.Rink, R. F. Irvine, Biochem. J. 235:869-877 (1986) and by J. E. Merritt,R. Jacob, T. J. Hallam, J. of Biol. Chem. 264:1522-1527 (1989).

On individual HL-60 cells, the transmembranal influx through"unselective cation channels" after ATP stimulation was measuredelectrophysiologically by the patch clamp technique. This influx isinhibited by Compound A (IC₅₀ : 7 nM).

These findings are evidence of the direct point of attack on isolatedcells. In particular, in the living organism, the granulocytes orleucocytes used in other tests migrate, after damage to brain cells(e.g. caused by stroke) into the damaged area, where they are activatedby calcium-mediated processes, decompose and thereby releasetissue-damaging mediators, including some chemotactic ones (e.g. PAF,leucotrienes, prostaglandins, FMLP etc.). Additional leucocytesattracted by chemotaxis increase the size of the damaged area. The testresults described above show that this vicious circle can be stopped bythe administration of the active substances described above. As a resultthe neurological damage is limited.

It has been demonstrated that conventional calcium antagonists (e.g.verapamil, nifedipine, diltiazem) do not inhibit the activation ofleucocytes.

Further tests with Compound A support the above finding:

The direct neuronal attack of Compound A was demonstrated on isolatedcortical and hippocampal neurone cells from foetal rat brains (preparedaccording to H. W. Muller and W. Seifert, Proc. Natl. Acad. Sci. USA 81:1248-1252, 1984; J. Neurosci. Res. 8: 195-204, 1982; and Muller andSeifert in "Methods for Serum Free Culture of Neuronal and LymphoidCells," p. 67-77, A. R. Liss Inc., 150 Fifth Ave., New York, N.Y. 10011,1984). In FURA₂ -charged single cells the concentration-time curves(calcium transient) of the cytoplasmic Ca²⁺ were recorded (method:modified from J. A. Connor, Proc. Natl. Acad. Sci. 83: 6179-6183, 1986).Both after mechanical lesion and after the administration of excitatoryamino acids (E.A.A., glutamate, cainate, quisqualate and NMDA) a sharpincrease in the cytoplasmic calcium concentrations was induced, whichcould be inhibited by Compound A in every case as a function of dosage(IC₅₀ at 3 μM).

The mechanism of activity of this inhibition was investigated both onneuronal cell cultures and also on human neutrophilic granulocytes andHL 60 cells and thrombocytes. It has been shown that Compound A inhibitsthe transmembranal flow of calcium into the cells stimulated by receptoragonists (e.g. EAA, FMLP, leucotriene, PAF, endotheline etc.). Thisinflux referred to by T. J. Hallam and T. J. Rink (Tips 10: 8-10, 1989)as "receptor mediated Ca²⁺ entry (RMCE)" cannot be inhibited byconventional calcium antagonists. Conventional calcium antagonistscannot prevent the leucocyte and thrombocyte activation since thesecells do not have any voltage-dependent Ca²⁺ channels. The blockade ofthe transmembranal calcium influx has been confirmed byelectrophysiological means (voltage clamp technique) on HL60 cells andneurone cells.

The inhibition of blood clotting or blood platelet aggregation accordingto the invention can be demonstrated by standard tests: on QUIN2-charged human blood platelets which have been stimulated with ADP,vasopressin, PGF2α, thrombin or serotonin, it can be demonstrated thatthe intracellular Ca-transient which results in platelet aggregation isinhibited by 3 μmol of Compound A.

In Patent Application EP-A-251 194 which has already been referred to,it is mentioned that in vitro tests on the smooth muscle (strips ofaorta; C. van Breemen, P. Aarenson, R. Lautzenheiser, K. Meisheri, Chest78: 157S-165S (1980); R. Casteels and G. Droogman, J. Physio. 317:263-279 (1981) have shown that the compounds of formula I are calciumantagonists with a new mechanism of activity. It is emphasised, to makematters clear, that the tests referred to demonstrate the cardiovasculareffect of the compounds. However, the test results obtained by them donot in any way make it obvious for the compounds of general formula I tohave an effect on inflammatory or neuronal cells.

The compounds may be given orally, parenterally or topically. Suitableforms include, for example, tablets, capsules, suppositories, solutions,syrups, emulsions, aerosols or dispersible powders. Tablets may beproduced, for example, by mixing the active substance or substances withknown excipients, e.g. inert diluents such as calcium carbonate, calciumphosphate or lactose, disintegrants such as corn starch or alginic acid,binders such as starch or gelatine, lubricants such as magnesiumstearate or talc and/or agents for obtaining delayed release, such ascarboxypolymethylene, carboxymethylcellulose, cellulose acetatephthalate or polyvinylacetate. The tablets may also consist of severallayers.

Coated tablets may be produced analogously by coating cores made in thesame way as the tablets with substances conventionally used for tabletcoatings, e.g. collidone or shellack, gum arabic, talc, titanium dioxideor sugar. In order to obtain delayed release or avoid incompatibilities,the core may also consist of several layers. Similarly, the tabletcoating may consist of several layers to achieve delayed release, whilstthe excipients mentioned for the tablets may be used.

Syrups containing the active substances or combinations of activesubstances according to the invention additionally contain a sweetenersuch as saccharin, cyclamate, glycerol or sugar as well as a flavourenhancer, e.g. a flavouring such as vanillin or orange extract. They mayalso contain suspension adjuvants or thickeners such as sodiumcarboxymethylcellulose, wetting agents, e.g. condensation products offatty alcohols with ethylene oxide or preservatives such asp-hydroxybenzoates.

Injectable solutions are produced in the usual way, e.g. by addingpreservatives such as p-hydroxybenzoates or stabilisers such as alkalimetal salts of ethylene diamine tetraacetic acid, and are thentransferred into injection vials or ampoules.

Capsules containing one or more active substances or combinations ofactive substances may be prepared for example by mixing the activesubstances with inert carriers such as lactose or sorbitol andencapsulating them in gelatine capsules.

Suitable suppositories may be produced for example by mixing withcarriers provided for this purpose, such as neutral fats orpolyethyleneglycol or derivatives thereof.

The compounds may be administered according to the invention by enteral,parenteral or topical route, advantageously in a quantity of 0.05 to 500mg per dose for adults. Preferably, 0.1 to 500 mg are used per dose fororal administration and 0.05 to 150 mg per dose for intravenousadministration.

Examples of formulations

    ______________________________________                                        Example 1: Tablets                                                            ______________________________________                                        Active substance according to invention                                                                40.0   mg                                            Lactose                  100.0  mg                                            Corn starch              50.0   mg                                            Colloidal silica         2.0    mg                                            Magnesium stearate       3.0    mg                                            Total                    200.0  mg                                            ______________________________________                                    

Method:

The active substance is mixed with some of the excipients and granulatedwith a solution of the soluble starch in water. After the granules havedried the remaining excipients are added and the mixture is compressedto form tablets.

    ______________________________________                                        Example 2: Coated tablets                                                     ______________________________________                                        Active substance according to invention                                                                20.0   mg                                            Lactose                  100.0  mg                                            Corn starch              65.0   mg                                            Colloidal silica         2.0    mg                                            Soluble starch           5.0    mg                                            Magnesium stearate       3.0    mg                                            Total                    195.0  mg                                            ______________________________________                                    

Method:

The active substance and excipients are compressed to form tablet coresas described in Example 1 and these are then coated with sugar, talc andgum arabic in the usual way.

    ______________________________________                                        Example 3: Suppositories                                                      ______________________________________                                        Active substance according to invention                                                                50.0   mg                                            Lactose                  250.0  mg                                            Suppository mass q.s. ad 1.7    g                                             ______________________________________                                    

Method:

The active substance and lactose are mixed together and the mixture isuniformly suspended in the molten suppository mass. The suspensions arepoured into cooled moulds to form suppositories weighing 1.7 g.

    ______________________________________                                        Example 4: Ampoules                                                           ______________________________________                                        Active substance according to the invention                                                            20.0   mg                                            Sodium chloride          5.0    mg                                            Twice distilled water q.s. ad                                                                          2.0    ml                                            ______________________________________                                    

Method:

The active substance and sodium chloride are dissolved in twicedistilled water and the solution is filtered into ampoules under sterileconditions.

    ______________________________________                                        Example 5: Ampoules                                                           Active substance according to invention                                                               10.0    mg                                            Sodium chloride         7.0     mg                                            Twice distilled water q.s. ad                                                                         1.0     mg                                            Example 6: Drops                                                              Active substance according to invention                                                               0.70    g                                             Methyl p-hydroxybenzoate                                                                              0.07    g                                             Propyl p-hydroxybenzoate                                                                              0.03    g                                             Demineralised water q.s. ad                                                                           100.00  ml                                            ______________________________________                                    

Method:

The active substance and preservatives are dissolved in demineralisedwater, the solution is filtered and transferred into vials each holding100 ml.

As has already been mentioned above, the compounds of formula Ie arenew. They can be prepared analogously to the processes described inEP-A-00 37 934 and EP-A-251 194:

a) by cyclising a phenylmalonic acid diamide of formula III ##STR96##

in the presence of a condensation agent

or

b) starting from carboxylic acids or carboxylic acid halides of formulaIV wherein Y represents OH or halogen, by reacting with the amine ofgeneral formula VI ##STR97## in the presence of a suitable condensationagent.

In the above formulae III and IV, is as defined for formula Ie.

Suitable condensation agents for process a) according to the inventioninclude numerous Lewis acids such as, e.g., phosphorus oxychloride,boron trifluoride, tin tetrachloride or titanium tetrachloride, as wellas strong inorganic acids such as sulphuric, fluorosulphonic,hydrofluoric or polyphosphoric acid. They are generally used in anexcess. Phosphorus oxychloride is preferred.

The reaction of cyclisation may be carried out in the presence orabsence of a solvent. All inert solvents are suitable provided that theyhave sufficient solubility for the reactants and have a high enoughboiling point. Examples include benzene, alkylbenzenes, chlorobenzenes,decalin, chloroform, methylene chloride, acetonitrile and the like. In avariant of the process the condensation agent, such as phosphorusoxychloride, is itself used as solvent.

No particular conditions apply to the reaction temperature. The reactionaccording to the invention can be carried out within a wide temperaturerange, preferably with warming or heating up to about the boiling pointof the solvent.

The reaction of amidation b) can theoretically be carried out under thesame conditions as reaction a). Examples of condensation agentsadditionally include carbodiimides such as cyclohexylcarbodiimide orcarbonyldiimidazole.

EXAMPLE

3,4-Dihydro-1-benzyl-6,7-dimethoxy-α-di-2-(2,3,4-trimethoxyphenyl)ethyl!aminocarbonyl-isoquinolinehydrochloride ##STR98## a)2-(3,4-dimethoxyphenyl)ethylaminocarbonyl-phenylacetic acid-N,N-di- 2(2,3,4-trimethoxyphenyl)-ethyl!amide

At ambient temperature, 9.0 g (55.5 mmol) of N,N'-carbonyldiimidazoleare stirred, in batches, into a solution of 18.0 g (52.4 mmol) ofmonoethyl-phenylmalonate-2-(3,4-dimethoxyphenyl)ethylamide in 150 ml ofanhydrous dimethylformamide. After 30 minutes, 18.0 g (44.3 mmol) of di-2-(2,3,4-trimethoxy-phenyl)ethylamine are added and the mixture isstirred for 30 minutes. The solvent is then distilled in vacuo, theresidue is taken up in 1.5 liters of CH₂ Cl₂ and extracted twice insuccession with 250 ml of water and 200 ml of 1N HCl. The organic phaseis dried over Na₂ SO₄, concentrated by evaporation, and the residue ispurified over a silica gel column (eluant: CH₂ Cl₂ /MeOH 100:2) andcrystallised from ethyl acetate/ether.

Yield: 35.5 g

b) 35.0 g (47.5 mmol) of amide (from Step a)) and 15 ml (164 mmol) ofphosphorus oxychloride are heated to boiling for 30 minutes in 150 ml ofanhydrous acetonitrile. After the reaction has ended (monitored by thinlayer chromatography) the solvent and any unused phosphorus oxychlorideare distilled off in vacuo. The residue is mixed with ice water, madealkaline with soda solution and extracted with about 1 liter of CH₂ Cl₂in batches. The organic phase is washed with water, dried over Na₂ SO₄and concentrated by evaporation. The residue is purified twice over asilica gel column (1st eluant: CH₂ Cl₂ :MeOH 100:2→100:4 ascending; 2ndeluant: CH₂ Cl₂ /ethyl acetate 1:1).

The hydrochloride is formed from the purified product (6.5 g) bydissolving in about 50 ml of ethanol and adding alcoholic hydrochloricacid. After evaporation and drying in a high vacuum at 50° C., 11.5 g ofthe desired product remain.

(Mp. 56°-64° C., amorphous)

Analogously to the Example, the Compounds of Table 12 can be prepared.

What is claimed is:
 1. A method of treating chronic inflammatoryprocesses in a warm-blooded animal which comprises administering to saidanimal a therapeutically effective amount of a carbocyclically andheterocyclically fused dihydropyridine of formula ##STR99## or thetautomeric form thereof of formula ##STR100## wherein X represents OR₁ ;NHR₂ ; NR₃ R₄ andR₁ represents hydrogen, C₁₋₄ -alkyl, C₁₋₄ -alkoxyalkylR₂ represents hydrogen; C₃₋₆ -alkenyl, C₃₋₆ -alkynyl; C₃₋₆ -cycloalkyl;C₃₋₆ -cycloalkenyl; straight-chained or branched C₁₋₆ -alkyl which mayif desired be mono- or polysubstituted with the substituents of groupsa) to c) listed below, which may be identical or different: a) halogen;cyano; hydroxy; mercapto; C₁₋₄ -alkoxy; C₁₋₄ -alkylthio; amino;mono-C₁₋₄ -alkylamino; di-C₁₋₄ -alkylamino (wherein the alkyl radicalmay be identical or different), phenoxy (wherein the phenyl ring may besubstituted as described as b) below) b) phenyl; optionally mono- orpolysubstituted, by identical or different substituents, by the groupshalogen, trifluoromethyl, C₁₋₄ -alkoxy, hydroxy, mercapto, C₁₋₄-alkylthio, C₁₋₄ -alkyl, amino, mono-C₁₋₄ -alkylamino, di-C₁₋₄-alkylamino (wherein the alkyl groups may be identical or different),C₂₋₃ -acylamino, C₂₋₃ -acyloxy and the group --O--(CH₂)_(n) --Ovicinally by bound to the phenyl system (wherein n=1 or 2) c) a 5- or6-membered saturated or wholly or partially unsaturated monocyclicheterocyclic group having up to 3 heteroatoms selected from the group N,O, S; and as a bicyclic heterocycle indole (whilst the above-mentionedheterocycles may be mono- or polysubstituted by C₁₋₄ -alkyl), C₃₋₆-cycloalkyl; C₅ - or C₆ -cycloalkenyl; C₂₋₃ -acyl; C₁₋₄ -alkylsulphonyl;or phenyl (which may in turn be substituted up to three times asdescribed in b));or R₂ represents phenyl, which may be substituted asdescribed in b) above; R₃ and R₄ independently of each other representC₁₋₄ -alkyl, which may if desired be phenyl-substituted, whilst thephenyl substituent may in turn be substituted as under b) hereinbefore;or R₃ and R₄ together with the nitrogen atom to which they are boundrepresent a wholly or partially saturated heterocyclic 5- or 6-memberedring (which may also contain up to 2 further heteroatoms from the groupN, O, S), whilst the heterocyclic group thus obtained may be substitutedby C₁₋₄ -alkyl, hydroxy or (CH₂)_(p) -R₅ (where p=0 or 1) and R₅represents a phenyl radical which is optionally substituted as under b)hereinbefore; A represents the fused ring systems ##STR101## wherein R₈represents hydrogen; C₁₋₄ -alkyl or C₁₋₄ -alkoxy R₆ and R₇ which may beidentical or different represent hydrogen; hydroxy; C₁₋₄ -alkyl; C₁₋₄-alkoxy; amino; methanesulphonylamino or R₆ and R₇ together represent--O--(CH₂)_(n) --O-- (where n=1 or 2) R₉ represents hydrogen; C₁₋₄-alkyl R₁₀ represents hydrogen or 2-phenyl-2-ethoxycarbonylacetyl; andthe salts thereof with physiologically acceptable acids, bases orcomplex-forming agents.
 2. The method as recited in claim 1 wherein thecarbocyclically and heterocyclically fused dihydropyridine is of formulaI or II,wherein X represents OR₁ NHR₂ ; NR₃ R₄ R₁ represents methyl orethyl R₂ represents hydrogen; straight-chained or branched unsubstitutedC₁₋₅ -alkyl; allyl; propargyl; C₃₋₆ -cycloalkyl; 3-chlorophenyl;2-methyl-3-chlorophenyl; or C₁₋₃ -alkyl, which is mono-substituted withone of the substituents of groups d) to f) listed hereinafter; d) cyano,hydroxy, methoxy, dimethylamino e) phenyl, 3,4-methylenedioxyphenyl,mono-, di- or trimethoxy substituted, phenyl, 3-methoxy-4-hydroxphenyl,3-hydroxy-4-methoxyphenyl, f) morpholino, pyridin-2-yl, indol-3-yl,furan-2-yl, thiphen-2-yl, pyridin-3-yl, pyridin-4-ylR₃ and R₄independently of each other represent methyl; ethyl; 3-cyanopropyl;benzyl; or 3,4,5-trimethoxyphenethyl or R₃ and R₄ together with thenitrogen atom to which they are bound represent morpholine;thiomorpholine; pyrrolidine; piperazine; 4-methylpiperazine;4-benzylpiperazine; or 4-(2-methoxyphenyl)piperazine; and A representsthe fused ring systems ##STR102## wherein R₈ represents hydrogen ormethoxyR₆ represents methoxy; hydroxy; hydrogen; amino; ormethanesulphonylamino R₇ represents hydrogen; methoxy; or hydroxy R₉represents methyl R₁₀ represents 2-phenyl-2-ethoxycarbonylacetyl; orhydrogen.
 3. The method as recited in claim 1 wherein thecarboxycyclically and heterocyclically fused dihydropyridine is offormula I or II, whereinA is the group ##STR103##
 4. The method asrecited in claim 1 wherein the carboxycyclically and heterocyclicallyfused dihydropyridine is of formula I or II, wherein R₈ is hydrogen andR₆ and R₇ independently of one another represent hydroxy or methoxy ortogether represent --O--CH₂ --O--, R₆ and R₇ preferably representingmethoxy.
 5. The method as recited in claim 1 wherein thecarboxycyclically and heterocyclically fused dihydropyridine is offormula I or II, wherein X is --NHR₂ or --NR₃ R₄.
 6. The method asrecited in claim 5 wherein the carboxycyclically and heterocyclicallyfused dihydropyridine is of formula I or II, wherein X is NHR₂, and R₂represents C₁₋₆ -alkyl.
 7. The method as recited in claim 6 wherein thecarboxycyclically and heterocyclically fused dihydropyridine is offormula I or II, wherein R₂ is C₁₋₄ -alkyl which is unsubstituted orsubstituted by hydroxy, phenyl, (which may be substituted by hydroxy,methoxy or --O--CH₂ --O--) or morpholino.
 8. The method as recited inclaim 1 wherein the carboxycyclically and heterocyclically fuseddihydropyridine is of the formula ##STR104## or a pharmaceuticallyacceptable salt thereof.
 9. The method as recited in claim 1 wherein thecarboxycyclically and heterocyclically fused dihydropyridine is of theformula ##STR105## or a pharmaceutically acceptable salt thereof.